Analysis of deoxynivalenol and deoxynivalenol-3-glucosides content in Canadian spring wheat cultivars inoculated with Fusarium graminearum

Contamination of wheat grains with Fusarium mycotoxins and their modified forms is an important issue in wheat industry. The objective of this study was to analyse the deoxynivalenol (DON) and deoxynivalenol-3-glucosides (D3G) content in Canadian spring wheat cultivars grown in two locations, inoculated with a mixture of 3-acetyldeoxynivalenol (3-ADON)-producing Fusarium graminearum strains and a mixture of 15-acetlyldeoxynivalenol (15-ADON)-producing F. graminearum strains. According to the analysis of variance, significant differences were observed among the cultivars for Fusarium head blight (FHB) disease index, Fusarium-damaged kernel percentage (%FDK), DON content and D3G content. When the effect of chemotype was considered, significant differences were observed for FHB disease index, FDK percentage and DON content. The D3G content and D3G/DON ratio were not significantly different between the chemotypes, except for D3G content at the Winnipeg location. The Pearson correlation coefficient between DON and D3G was 0.84 and 0.77 at Winnipeg and Carman respectively. The highest D3G/DON ratio was observed in cultivars Carberry (44%) in Carman and CDC Kernen (63.8%) in Winnipeg. The susceptible cultivars showed lower D3G/DON ratio compared with the cultivars rated as moderately resistant and intermediate. The current study indicated that Canadian spring cultivars produce D3G upon Fusarium infection.     

Less Fusarium infestation and mycotoxin contamination in organic than in conventional cereals

A total of 602 samples of cereals, consisting of organically and conventionally produced barley, oats and wheat, were collected at harvest during 2002–2004 in Norway. Organic and conventional cereals were sampled in comparable numbers regarding cereal species, localisation and harvest time, and analysed for Fusarium mould and mycotoxins. Fusarium infestation and mycotoxin content were dependent on cereal species and varied year-by-year. However, in all cereal species, Fusarium infestation and levels of important mycotoxins were significantly lower when grown organically than conventionally. Concerning the most toxic trichothecenes, HT-2 and T-2 toxin, lower concentrations were found in organic oats and barley. Wheat was not contaminated by HT-2 and T-2, but lower concentrations of deoxynivalenol (DON) and moniliformin (MON) were found when organically produced. For mycotoxins considered to constitute the main risk to humans and animals in Norwegian cereals, i.e. HT-2 in oats and DON in oats and wheat, the median figures (mean levels in brackets) were as follows: HT-2 in organic and conventional oats were <20 (80) and 62 (117) µg/kg, DON in organic and conventional oats were 24 (114) and 36 (426) µg/kg, and DON in organic and conventional wheat were 29 (86) and 51 (170) µg/kg, respectively. Concentrations of HT-2 and T-2 in the samples were strongly correlated (r = 0.94). Other mycotoxins did not show a significant correlation to each other. Both HT-2 and T-2 concentrations were significantly correlated with infestation of F. langsethiae (r = 0.65 and r = 0.60, respectively). Concentrations of DON were significantly correlated with F. graminearum infestation (r = 0.61). Furthermore, nivalenol (NIV) was significantly correlated with infestation of F. poae (r = 0.55) and MON with F. avenaceum (r = 0.37). As lower Fusarium infestation and mycotoxin levels were found in organic cereals, factors related to agricultural practice may reduce the risk of contamination with Fusarium mycotoxins. Studies of these issues will be presented separately.     

Relationship between Fusarium spp. diversity and mycotoxin contents of mature grains in southern Belgium

Over a 4-year period (2010–13), a survey aiming at determining the occurrence of Fusarium spp. and their relations to mycotoxins in mature grains took place in southern Belgium. The most prevalent species were F. graminearum, F. avenaceum, F. poae and F. culmorum, with large variations between years and locations. An even proportion of mating type found for F. avenaceum, F. culmorum, F. cerealis and F. tricinctum is usually a sign of ongoing sexual recombination. In contrast, an unbalanced proportion of mating type was found for F. poae and no MAT1-2 allele was present in the F. langsethiae population. Genetic chemotyping indicates a majority of deoxynivalenol (DON)-producing strains in F. culmorum (78%, all 3-ADON producers) and F. graminearum (95%, mostly 15-ADON producers), while all F. cerealis strains belong to the nivalenol (NIV) chemotype. Between 2011 and 2013, DON, NIV, enniatins (ENNs) and moniliformin (MON) were found in each field in various concentrations. By comparison, beauvericin (BEA) was scarcely detected and T-2 toxin, zearalenone and α- and β-zearalenols were never detected. Principal component analysis revealed correlations of DON with F. graminearum, ENNs and MON with F. avenaceum and NIV with F. culmorum, F. cerealis and F. poae. BEA was associated with the presence of F. tricinctum and, to a lesser extent, with the presence of F. poae. The use of genetic chemotype data revealed that DON concentrations were mostly influenced by DON-producing strains of F. graminearum and F. culmorum, whereas the concentrations of NIV were influenced by the number of NIV-producing strains of both species added to the number of F. cerealis and F. poae strains. This study emphasises the need to pay attention to less-studied Fusarium spp. for future Fusarium head blight management strategies, as they commonly co-occur in the field and are associated with a broad spectrum of mycotoxins.     

Mycoflora and deoxynivalenol in whole wheat grains (Triticum aestivum L.) from Southern Brazil

The fungal species Fusarium graminearum is related to deoxynivalenol (DON) formation. The aim of this study was to evaluate mycoflora and DON occurrence in 53 whole wheat grain samples collected in Southern Brazil during the 2012 crop. Wheat grains showed adequate values of water activity ranging from 0.48 to 0.72, within the required limits of moisture content, ranging from 9.1% to 13.9%. In addition, low counts of fungal colonies, ranging from 10 to 8.2 × 10², were found. For Fusarium genera, there was predominance of Fusarium verticillioides (34%) and F graminearum (30.2%). For Aspergillus species, 37.7% of Aspergillus flavus was determined. Regarding the Penicillium species, Penicillium digitatum (49%) was the most found species. DON was detected in 47.2% (25 out of 53) of the samples analysed, with levels ranging from 243.7 to 2281.3 µg kg^?1 (mean: 641.9 µg kg^?1).     

Mycotoxin profiles in the grain of Triticum monococcum,Triticum dicoccum and Triticum spelta after head infection with Fusarium culmorum

BACKGROUND: The aim of study was to investigate mycotoxin profiles in the grain of spring lines of Triticum monococcum (12 lines), T. dicoccum (13 lines) and T. spelta (five lines), in comparison to the T. aestivum cultivar Sumai‐3 which is resistant to Fusarium head blight. The grain was obtained from control heads and heads artificially inoculated in the field with Fusarium culmorum. Mycotoxins were determined by LC‐MS/MS. RESULTS: A total of 11 toxins were identified in control grain samples. Deoxynivalenol (DON) concentrations exceeded 0.5 mg kg^?1 in only three samples of T. monococcum grain and in one sample of T. dicoccum grain. Inoculation with F. culmorum resulted in a substantial increase in the concentrations of DON (to 63 mg kg^?1) in the T. monococcum and DON‐3‐glucoside (to 5.6 mg kg^?1) in the T. dicoccum. Inoculation contributed to a drop in tentoxin levels (by 57% in T. spelta) and to an increase of cyclodepsipeptide concentrations (in particular enniatins B and B₁) being two‐fold (T. monococcum) to four‐fold (T. dicoccum) higher on average than in control samples. The Sumai‐3 responded to inoculation with nearly a two‐fold drop in the levels of the cyclodepsipeptides. CONCLUSION: The results of a discriminant analysis for all identified toxins indicate that einkorn, emmer and spelt differ significantly with regard to the mycotoxin profiles of their grain. Copyright © 2009 Society of Chemical Industry     

Contaminants in organically and conventionally produced winter wheat (Triticum aestivum) in Belgium

A database has been compiled with the levels of important contaminants (mycotoxins, heavy metals and pesticides) measured from 2002 to 2005 in winter wheat (Triticum aestivum) grown in Belgium according to the organic and conventional farming systems. Assuming no further change in contaminant levels during cereal processing and during the preparation of foodstuffs, conservative intakes are estimated for the consumers of cereal-based products such as flour, bread, breakfast cereals, dough and pastry. The results show that for the consumer of organic foodstuffs, estimated daily intakes are 0.56 µg deoxynivalenol (DON), 0.03 µg zearalenone (ZEA), 0.19 µg Cd, 0.28 µg Pb and 0.0006 µg Hg kg^-1 body weight, taking into account the average contaminant levels in unprocessed grains and the average cereal products consumptions in Belgium. For the consumers of conventional foodstuffs, the corresponding estimated daily intakes are 0.99 µg DON, 0.06 µg ZEA, 0.17 µg Cd, 0.12 µg Pb and 0.0007 µg Hg kg^-1 body weight. In addition, it appears that for the consumers of conventional products, intakes of some post-harvest insecticides have to be taken into account (0.11 µg chlorpyriphos-methyl, 0.2 µg dichlorvos and 0.24 µg pirimiphos-methyl kg^-1 bw). When expressed as a percentage of the tolerable/acceptable daily intake (TDI/ADI), it seems that the corresponding estimated (conservative) intakes are the highest for DON (56% for organic and 99% for conventional cereal products), ZEA (16% for organic and 32% for conventional cereal products), and Cd (19% for organic and 17% for conventional cereal products), all other estimated intakes of contaminants (including pesticides) being lower than 10% of the TDI/ADI.     

Assessment of Fusarium infection in wheat heads using a quantitative polymerase chain reaction (qPCR) assay

The accuracy of a quantitative polymerase chain reaction assay in quantifying the DNA of trichothecene-producing F. culmorum and F. graminearum within harvested wheat grains and head tissue was evaluated in comparison with incidences of infected kernels and deoxynivalenol levels. In a first experiment, six durum and bread wheat varieties were grown in randomized plots for a 2-year period, and inoculated with Fusarium macroconidia at six growth stages between heading and dough ripening, to obtain a wide range of Fusarium head blight incidences. There was a close relationship between fungal DNA and the amount of deoxynivalenol, and this relationship was consistent over Fusarium species, wheat species and varieties, and over a wide range of Fusarium head blight infection. In a second experiment potted wheat plants were grown under environmentally controlled conditions and inoculated with the two Fusarium species at full flowering; head samples were collected before inoculation and after 6 h to 12 days, and processed by the quantitative polymerase chain reaction assay. This assay made it possible to detect the dynamic of fungal invasion in planta after infection had occurred, and to single out the presence of infection before the onset of the disease symptoms: A robust detection of the infection occurred within 18-24 h for F. culmorum, and within 2-9 days for F. graminearum.     

Fusarium head blight and associated mycotoxin occurrence on winter wheat in Luxembourg in 2007/2008

Fusarium head blight (FHB) is among the major causes of reduced quality in winter wheat and its products. In addition, the causal fungi produce a variety of toxins. A relatively high FHB infection rate in winter wheat was observed in 2007 and 2008 in Luxembourg. A fusariotoxin survey was carried out in 17 different geographical locations. Three groups of Fusarium mycotoxins (trichothecenes A and B and zearalenone) were analysed by a multi-detection HPLC–MS/MS method. Fusarium strains were also investigated by morphological and molecular methods. In addition, questionnaires relating to cultural practices were sent to the farmers managing the 17 fields investigated. FHB prevalence ranged from 0.3 to 65.8% (mean: 8.5%) in 2007 and from 0 to 24.5% (mean: 8.3%) in 2008. Results of morphological and molecular identification showed that the most common species isolated from diseased wheat spikes was F. graminearum (33.1%), followed by F. avenaceum (20.3%) and F. poae (17.8%). The chemical analysis revealed that 75% of the investigated fields were contaminated by deoxynivalenol (DON, range 0–8111 µg/kg). The preceding crop was highly and significantly correlated to the number of grains infected and had a significant impact on disease prevalence (p = 0.025 and 0.017, respectively, Fisher's F-test). A trend was found for maize as the preceding crop (p = 0.084, Tukey's test) to predict the amount of DON in the fields. This is the first report on the occurrence of DON and ZON in naturally infected wheat grains sampled from Luxembourg.     

Distribution of toxigenic Fusarium spp. and mycotoxin production in milling fractions of durum wheat

A reliable and sensitive PCR assay to specifically detect trichothecene-producing Fusarium spp. in milling fractions and kernel tissue of naturally infected durum wheat is reported. Assays were based on a combination of primers derived from the trichodiene synthase and the β-tubulin genes. The occurrence of toxigenic Fusarium spp. in semolina and wheat tissue (grain ends, crease, pericarp, aleurone layer, germ and albumen) was detected, even for a weakly contaminated wheat sample. Penetration of toxigenic Fusarium spp. into the interior of durum wheat kernel was demonstrated for the Nefer variety, indicating that none of the tissue structures within the wheat kernel acted as an effective barrier to fungal invasion. Moreover, after inoculation by toxigenic Fusarium strains, semolina was shown to allow high yields of trichothecenes, while bran was demonstrated to contain biochemical inhibitors able to significantly reduce trichothecene production. These results will be useful in improving breeding strategies to control trichothecene contamination of durum wheat kernels.     

Relationship between free glutathione content and quality assessment parameters of wheat cultivars (Triticum aestivum L.)

Freshly milled flour samples from ten cultivars of bread wheat were analysed for free reduced glutathione (GSH) and oxidised glutathione (GSSG) by an enzymic method. Dough rheological properties for each flour were assessed by standard Brabender techniques. Flour quality differences were also measured by Pelshenke fermentation and Zeleny sedimentation techniques. Reduced glutathione levels were negatively correlated with oxidised glutathione levels, Farinogram mixing tolerance, Extensogram resistance, Pelshenke time and Zeleny volume. The ratios of oxidised to reduced glutathione (GSSG: GSH) were positively correlated with all dough and flour quality parameters, except Extensogram extensibility. It is concluded that there are differences in the endogenous contents of reduced and oxidised glutathione between flour samples from ten cultivars of wheat. These differences may contribute significantly to variations in the quality assessment parameters established between cultivars in this study.     

Effect of fungicides on the development of Fusarium head blight,yield and deoxynivalenol accumulation in wheat inoculated under field conditions with Fusarium graminearum and Fusarium culmorum

The effect of different fungicide treatments on Fusarium head blight (FHB) development, grain yields and deoxynivalenol (DON) accumulation in wheat was evaluated after artificial inoculation under field conditions with a mixture of Fusarium graminearum and F culmorum. The trials were carried out using commercially available products on five different cultivars of soft and durum wheat (Serio, Genio, Bracco, Duilio and Orobel) in two separate experimental fields located in the North of Italy. Treatments with Tiptor^® S (cyproconazole plus prochloraz) and a mixture of Horizon^® 250 EW (tebuconazole) plus Amistar^® 250 SC (azoxystrobin) significantly reduced the FHB disease severity (by between 25 and 77%) and DON content (by between 32 and 89%) in the grain as compared with the inoculated control. Yields (tonne ha^?1) and thousand grain weight (TGW) were higher in plots subjected to fungicide treatments. Tetraconazole (Eminent^® 40 EW) showed a markedly reduced effectiveness compared with the other treatments. Regression analysis showed a strong correlation of disease severity with DON levels (positive correlation) and with yields or TGW (negative correlation) for individual cultivars and locations. Fusarium graminearum, F culmorum and F poae were the species most commonly isolated from all trials, including inoculated and non‐inoculated control plots. Copyright © 2004 Society of Chemical Industry     

Investigations on Fusarium spp. and their mycotoxins causing Fusarium ear rot of maize in Kosovo

After wheat, maize (Zea mays L.) is the second most important cereal crop in Kosovo and a major component of animal feed. The purpose of this study was to analyse the incidence and identity of the Fusarium species isolated from naturally infected maize kernels in Kosovo in 2009 and 2010, as well as the mycotoxin contamination. The disease incidence of Fusarium ear rot (from 0.7% to 40% diseased ears) on maize in Kosovo is high. The most frequently Fusarium spp. identified on maize kernels were Fusarium subglutinans, F. verticillioides/F. proliferatum and F. graminearum. Maize kernel samples were analysed by LC-MS/MS and found to be contaminated with deoxynivalenol (DON), DON-3-glucoside, 3-acetyl-DON, 15-acetyl-DON, zearalenone, zearalenone-14-sulphate, moniliformin, fumonisin B₁ and fumonisin B₂. This is the first report on the incidence and identification of Fusarium species isolated from naturally infected maize as well as the mycotoxin contamination in Kosovo.     

Effect of atmospheric cold plasma on the functional properties of whole wheat (Triticum aestivum L.) grain and wheat flour

Atmospheric cold plasma (ACP) has emerged as a novel processing technology, with demonstrated efficiencies in microbial inactivation. However, studies on the effects of ACP and potential to modify the functional properties of foods are sparse. The objective of this study is to determine the effect of ACP on physico-chemical and functional properties of wheat flour. In this study, both whole wheat grains and wheat flour were subjected to a dielectric barrier discharge (DBD) contained plasma reactor for a range of treatment times (5–30 min) at 80 kV. Plasma treatment increased the flour hydration properties of wheat flour. Rapid visco-analyser results showed an increase in pasting and the final viscosities of wheat flour. The decrease in both endothermic enthalpies and crystallinity was attributed to the depolymerization of starch and plasma-induced changes. Overall DBD-ACP treatment can be tailored to develop a plasma process with potential to improve functionality of wheat flour.     

Varietal differences in amino acid composition of wheat (Triticum aestivum L) grain

The differences in the amino acid composition of the developing grain between the three winter wheat (Triticum aestivum L) varieties Splendeur, Hobbit and Maris Huntsman were analysed in a growth-room experiment. At 15 days after anthesis, Maris Huntsman, the variety with the fastest leaf senescence, had a larger proportion of glutamate and smaller levels of threonine, glycine, alanine, valine, methionine and isoleucine in the total amino acid fraction; at later dates these differences were in general less pronounced, particularly compared with Splendeur. This suggests that, in the variety with the fastest leaf senescence, rapid gliadin synthesis in the grain occurs earlier. At any given percentage of nitrogen in the mature grain, Hobbit had a smaller proportion of glutamate and greater proportions of lysine, arginine and alanine; Splendeur, the only bread-making variety, had smaller levels of aspartate and lysine; and Maris Huntsman had the greatest relative amount of phenylalanine. The possible differences among the varieties in the relative proportion of some protein fractions are discussed. Until 25 days after anthesis Maris Huntsman had smaller relative proportions of free alanine, valine, leucine, isoleocine and threonine, in particular when compared with Splendeur. In Maris Huntsman, the relative amount of aspartate increased from 15 days after anthesis onwards, whereas in Hobbit it only did so in mature grains.     

Trichothecene and beauvericin mycotoxin production and genetic variability in Fusarium poae isolated from wheat kernels from northern Italy

The importance and widespread incidence of Fusarium poae as a natural contaminant of wheat in different climatic areas warrants investigation into the genetic diversity and toxin profile of a northern Italy population. Eighty-one strains of F. poae isolated from durum wheat kernels, identified by species-specific polymerase chain reaction and translation elongation factor-1α gene sequence analysis, were genetically characterized by the amplified fragment length polymorphism (AFLP) technique and analysed by high-pressure liquid chromatography for their ability to produce the beauvericin (BEA) and trichothecene mycotoxins. A high level of variability was observed by using AFLP analyses, with the lowest level of genetic similarity among the strains being approximately 61%. Most of the strains, 95%, produced BEA at <2655 µg g^?1; 88% produced the trichothecene nivalenol at <865 µg g^?1 and 76% produced the trichothecene fusarenon-X at <167 µg g^?1. These data show that F. poae can produce high amounts of BEA together with trichothecenes, and can represent a high potential mycotoxin risk in Italy for wheat colonized by this species.     

Influence of nitrogen fertilizer application rate on winter wheat (Triticum aestivum L.) flour quality and Chinese noodle quality

BACKGROUND: Understanding wheat flour noodle quality responses to N management will improve the quality of recommendations made to growers for specific end‐uses. Two winter wheat cultivars, with six N rates, planted during the course of a 2‐year experiment were used to determine the effect of N application rate on Chinese white noodle quality. RESULTS: Wheat flour protein content, development time (DT) and stability time (ST) increased with N application rate, maximal at 360 kg N ha^?1 and decreasing thereafter. When the N fertilizer application rate changed from 0 to 270 or 360 kg N ha^?1, redness (a*) and yellowness (b*) of both flour and noodle increased, while brightness (L*) decreased. The hardness and chewiness of cooked noodles improved at lower N application rate and degenerated at higher N rate. CONCLUSION: A low rate of N fertilizer application (from 0 to 270 kg N ha^?1) improved flour quality. Flour protein content and protein quality parameters displayed a significant negative correlation with brightness (L*) values. Generally, high flour protein content and protein quality, as well as good noodle texture, could be achieved by topdressing N fertilizer (in the range 270–360 kg N ha^?1). Copyright © 2009 Society of Chemical Industry